| pubmed-article:8532685 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C0023621 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C0521009 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C0597357 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C1514562 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C1883221 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C1883204 | lld:lifeskim |
| pubmed-article:8532685 | lifeskim:mentions | umls-concept:C1880389 | lld:lifeskim |
| pubmed-article:8532685 | pubmed:issue | 6 | lld:pubmed |
| pubmed-article:8532685 | pubmed:dateCreated | 1996-2-1 | lld:pubmed |
| pubmed-article:8532685 | pubmed:abstractText | The construction and characterization of a combinatorial library of a solvent-exposed surface of an alpha-helical domain derived from a bacterial receptor is described. Using a novel solid-phase approach, the library was assembled in a directed and successive manner utilizing single-stranded oligonucleotides containing multiple random substitutions for the variegated segments of the gene fragment. The simultaneous substitution of 13 residues to all 20 possible amino acids was carried out in a region spanning 81 nucleotides. The randomization was made in codons for amino acids that were modelled to be solvent accessible at a surface made up from two of the three alpha-helices of a monovalent Fc-binding domain of staphylococcal protein A. After cloning of the PCR-amplified library into a phagemid vector adapted for phage display of the mutants, DNA sequencing analysis suggested a random distribution of codons in the mutagenized positions. Four members of the library with multiple substitutions were produced in Escherichia coli as fusions to an albumin-binding affinity tag derived from streptococcal protein G. The fusion proteins were purified by human serum albumin affinity chromatography and subsequently characterized by SDS-electrophoresis, CD spectroscopy and biosensor analysis. The analyses showed that the mutant protein A derivatives could all be secreted as soluble full-length proteins. Furthermore, the CD analysis showed that all mutants, except one with a proline introduced into helix 2, have secondary structures in close agreement with the wild-type domain. These results proved that members of this alpha-helical receptor library with multiple substitutions in the solvent-exposed surface remain stable and soluble in E. coli.(ABSTRACT TRUNCATED AT 250 WORDS) | lld:pubmed |
| pubmed-article:8532685 | pubmed:language | eng | lld:pubmed |
| pubmed-article:8532685 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:8532685 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:8532685 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:8532685 | pubmed:month | Jun | lld:pubmed |
| pubmed-article:8532685 | pubmed:issn | 0269-2139 | lld:pubmed |
| pubmed-article:8532685 | pubmed:author | pubmed-author:NilssonBB | lld:pubmed |
| pubmed-article:8532685 | pubmed:author | pubmed-author:NilssonJJ | lld:pubmed |
| pubmed-article:8532685 | pubmed:author | pubmed-author:UhlénMM | lld:pubmed |
| pubmed-article:8532685 | pubmed:author | pubmed-author:NygrenP APA | lld:pubmed |
| pubmed-article:8532685 | pubmed:author | pubmed-author:PaulB NBN | lld:pubmed |
| pubmed-article:8532685 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:8532685 | pubmed:volume | 8 | lld:pubmed |
| pubmed-article:8532685 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:8532685 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:8532685 | pubmed:pagination | 601-8 | lld:pubmed |
| pubmed-article:8532685 | pubmed:dateRevised | 2006-11-15 | lld:pubmed |
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| pubmed-article:8532685 | pubmed:meshHeading | pubmed-meshheading:8532685-... | lld:pubmed |
| pubmed-article:8532685 | pubmed:year | 1995 | lld:pubmed |
| pubmed-article:8532685 | pubmed:articleTitle | A combinatorial library of an alpha-helical bacterial receptor domain. | lld:pubmed |
| pubmed-article:8532685 | pubmed:affiliation | Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden. | lld:pubmed |
| pubmed-article:8532685 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:8532685 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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